Microbial composition of goat buck's ejaculates is modified by the process of preparing and storing refrigerated semen doses

Ejaculates present their own microbiota, and a link between ejaculates' microbiota and sperm quality and fertility exists. With the development of artificial insemination in animal breeding, ejaculates must be manipulated by diluting them with extenders and storing them at temperatures below body te...

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Main Authors: Mocé, Maria Lorena, Esteve, Inés Carolina, Gómez, Ernesto A., Pérez-Fuentes, Sara, Mocé, Eva
Format: article
Language:Inglés
Published: Elsevier 2023
Subjects:
Online Access:https://hdl.handle.net/20.500.11939/8667
https://www.sciencedirect.com/science/article/abs/pii/S0093691X23002212
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author Mocé, Maria Lorena
Esteve, Inés Carolina
Gómez, Ernesto A.
Pérez-Fuentes, Sara
Mocé, Eva
author_browse Esteve, Inés Carolina
Gómez, Ernesto A.
Mocé, Eva
Mocé, Maria Lorena
Pérez-Fuentes, Sara
author_facet Mocé, Maria Lorena
Esteve, Inés Carolina
Gómez, Ernesto A.
Pérez-Fuentes, Sara
Mocé, Eva
author_sort Mocé, Maria Lorena
collection ReDivia
description Ejaculates present their own microbiota, and a link between ejaculates' microbiota and sperm quality and fertility exists. With the development of artificial insemination in animal breeding, ejaculates must be manipulated by diluting them with extenders and storing them at temperatures below body temperature. The effects that these processes have on the original semen microbiota have never been studied. This study explores the effects of the protocol for preparing refrigerated goat buck semen doses and storing on seminal microbiota. Semen from six adult goat bucks of the Murciano-Granadina breed (24 ejaculates) was used, cooled to 4 °C in a skimmed milk-based extender, and stored at this temperature for 24 h. Samples were taken in different steps: in the raw ejaculates (ejaculates), after dilution with the refrigeration extender (diluted), immediately after reaching 4 °C (chilled 0 h) and the samples refrigerated at 4 °C and stored at this temperature for 24 h (chilled 24 h). Sperm quality (motility and integrity of plasma and acrosomal membrane, and mitochondrial functionality) was also evaluated. Bacterial 16S rRNA sequencing was used to study the seminal microbiota. Our results indicated that both refrigeration and storage at 4 °C negatively affected sperm quality parameters. Preparing semen doses and their subsequent conservation caused a significant change in the bacterial community structure. Raw ejaculates showed a lower Pielou's evenness index than the other samples (diluted, chilled 0 h and chilled 24 h). Ejaculates also had a lower Shannon's diversity index (3.44) than the diluted semen (4.17) and the semen chilled for 24 h (4.43). Regarding beta diversity, significant differences were detected between ejaculates and the other treatments. Differences were also found in unweighted UniFrac distances between the semen chilled for 0 h and that chilled for 24 h. At the genus level, marked effects of preparing doses and their subsequent conservation were also evident: 199 genera that were absent in ejaculates were found in the semen chilled and stored for 24 h; 177 genera that were present in ejaculates disappeared after 24-h refrigeration. In conclusion, the extender and protocol for preparing refrigerated goat buck semen doses considerably modify microbial ejaculate composition.
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spelling ReDivia86672025-04-25T14:49:14Z Microbial composition of goat buck's ejaculates is modified by the process of preparing and storing refrigerated semen doses Mocé, Maria Lorena Esteve, Inés Carolina Gómez, Ernesto A. Pérez-Fuentes, Sara Mocé, Eva Goat bucks Microbial composition Ejaculates Sperm quality Murciano-Granadina Sperm metabolism L01 Animal husbandry L70 Veterinary science and hygiene L53 Animal physiology - Reproduction Semen preservation Refrigerated storage Fertility Microbiota Ejaculates present their own microbiota, and a link between ejaculates' microbiota and sperm quality and fertility exists. With the development of artificial insemination in animal breeding, ejaculates must be manipulated by diluting them with extenders and storing them at temperatures below body temperature. The effects that these processes have on the original semen microbiota have never been studied. This study explores the effects of the protocol for preparing refrigerated goat buck semen doses and storing on seminal microbiota. Semen from six adult goat bucks of the Murciano-Granadina breed (24 ejaculates) was used, cooled to 4 °C in a skimmed milk-based extender, and stored at this temperature for 24 h. Samples were taken in different steps: in the raw ejaculates (ejaculates), after dilution with the refrigeration extender (diluted), immediately after reaching 4 °C (chilled 0 h) and the samples refrigerated at 4 °C and stored at this temperature for 24 h (chilled 24 h). Sperm quality (motility and integrity of plasma and acrosomal membrane, and mitochondrial functionality) was also evaluated. Bacterial 16S rRNA sequencing was used to study the seminal microbiota. Our results indicated that both refrigeration and storage at 4 °C negatively affected sperm quality parameters. Preparing semen doses and their subsequent conservation caused a significant change in the bacterial community structure. Raw ejaculates showed a lower Pielou's evenness index than the other samples (diluted, chilled 0 h and chilled 24 h). Ejaculates also had a lower Shannon's diversity index (3.44) than the diluted semen (4.17) and the semen chilled for 24 h (4.43). Regarding beta diversity, significant differences were detected between ejaculates and the other treatments. Differences were also found in unweighted UniFrac distances between the semen chilled for 0 h and that chilled for 24 h. At the genus level, marked effects of preparing doses and their subsequent conservation were also evident: 199 genera that were absent in ejaculates were found in the semen chilled and stored for 24 h; 177 genera that were present in ejaculates disappeared after 24-h refrigeration. In conclusion, the extender and protocol for preparing refrigerated goat buck semen doses considerably modify microbial ejaculate composition. 2023-06-26T07:58:39Z 2023-06-26T07:58:39Z 2023 article publishedVersion Mocé, M. L., Esteve, I. C., Gómez, E. A., Pérez-Fuentes, S. & Mocé, E. (2023). Microbial composition of goat buck's ejaculates is modified by the process of preparing and storing refrigerated semen doses. Theriogenology, Published online [17 june 2023]. 0093-691X (print ISSN) 1879-3231 (online ISSN) https://hdl.handle.net/20.500.11939/8667 10.1016/j.theriogenology.2023.06.013 https://www.sciencedirect.com/science/article/abs/pii/S0093691X23002212 en This work was supported by the Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA) and was cofinanced by FEDER funds (grant number RTA2017-00049-C02-01), GVA-IVIA (grant number 51906) and AMURVAL (contract number 71714), and by the Universidad Cardenal Herrera-CEU, CEU Universities, grant number INDI21/40 and INDI22/38 info:eu-repo/grantAgreement/AEI/Programa Estatal de I+D+i Orientada a los Retos de la Sociedad/RTA2017-00049-C02-01 info:eu-repo/grantAgreement/ERDF/POCV 2014-2020/51906 Attribution-NonCommercial-NoDerivatives 4.0 Internacional http://creativecommons.org/licenses/by-nc-nd/4.0/ openAccess Elsevier electronico
spellingShingle Goat bucks
Microbial composition
Ejaculates
Sperm quality
Murciano-Granadina
Sperm metabolism
L01 Animal husbandry
L70 Veterinary science and hygiene
L53 Animal physiology - Reproduction
Semen preservation
Refrigerated storage
Fertility
Microbiota
Mocé, Maria Lorena
Esteve, Inés Carolina
Gómez, Ernesto A.
Pérez-Fuentes, Sara
Mocé, Eva
Microbial composition of goat buck's ejaculates is modified by the process of preparing and storing refrigerated semen doses
title Microbial composition of goat buck's ejaculates is modified by the process of preparing and storing refrigerated semen doses
title_full Microbial composition of goat buck's ejaculates is modified by the process of preparing and storing refrigerated semen doses
title_fullStr Microbial composition of goat buck's ejaculates is modified by the process of preparing and storing refrigerated semen doses
title_full_unstemmed Microbial composition of goat buck's ejaculates is modified by the process of preparing and storing refrigerated semen doses
title_short Microbial composition of goat buck's ejaculates is modified by the process of preparing and storing refrigerated semen doses
title_sort microbial composition of goat buck s ejaculates is modified by the process of preparing and storing refrigerated semen doses
topic Goat bucks
Microbial composition
Ejaculates
Sperm quality
Murciano-Granadina
Sperm metabolism
L01 Animal husbandry
L70 Veterinary science and hygiene
L53 Animal physiology - Reproduction
Semen preservation
Refrigerated storage
Fertility
Microbiota
url https://hdl.handle.net/20.500.11939/8667
https://www.sciencedirect.com/science/article/abs/pii/S0093691X23002212
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